大鼠(RAT),學(xué)名Rattus norvegicus,在生物分類學(xué)上屬脊椎動物門、哺乳動物綱、嚙齒目、鼠科、家鼠屬、褐家鼠種。實(shí)驗(yàn)室也常會看到大鼠的影子,大鼠在藥物學(xué)、代謝性疾病研究、腫瘤研究遺傳學(xué)等多個研究領(lǐng)域做出了很大的貢獻(xiàn)。此次試驗(yàn)的目的是研磨大鼠血管,提取核酸。
實(shí)驗(yàn)材料和器具:
樣品:大鼠血管
工具:凈信多樣品組織研磨儀(Tissuelyser-24)
耗材:離心管、研磨珠、液氮、剪刀、鑷子等
實(shí)驗(yàn)步驟:
1、準(zhǔn)備離心管,研磨珠若干;
2、首先將適配器置于裝有液氮的容器內(nèi),液氮容量沒過適配器即可,冷凍幾分鐘,取出時戴手套或使用鑷子夾取避免液氮凍傷;
3、從冰箱內(nèi)取大鼠血管組織剪成小塊與研磨珠一同放入離心管中,將蓋子蓋緊;
4、將適配器放入研磨儀中固定,然后在將離心管放置適合器內(nèi)。樣品盡量對稱放置,以保證研磨時的受力平衡,旋緊垂直定位螺旋,將安全鎖扣緊。設(shè)置參數(shù),程序結(jié)束后即可得到破碎粉末;
5、研磨完畢后請關(guān)閉電源,將安全鎖歸位;
6、完成后續(xù)核酸提取步驟。
注意事項(xiàng):
1、每支離心管樣品量一般不超過放心管體積的三分之二,樣品量過多會影響研磨效果。
2、樣品盡量對稱放置,以保證研磨時的受力平衡。
3、適配器采用半導(dǎo)體材質(zhì)需要提前放入液氮中浸泡幾分鐘預(yù)冷。
實(shí)驗(yàn)過程、結(jié)果圖示:
實(shí)用儀器介紹:
上海凈信?的Tiss組織研磨儀是一種特殊的、快速的、高效率的、多試管的一致系統(tǒng)。它能將任何來源(包括土壤、植物和動物的組織/器官、細(xì)菌、酵母、真菌、孢子、古生物標(biāo)本等)的原始DNA、RNA和蛋白質(zhì)進(jìn)行提取和純化。
采用了特殊的垂直上下一體震動模式,通過研磨珠(氧化鋯、鋼珠、玻璃珠、陶瓷珠)的高頻往復(fù)振動、撞擊、剪切。快速的實(shí)現(xiàn)目的。使研磨的樣品具有更加充分、更均勻、樣品重復(fù)性更好、樣品之間沒有交叉污染。
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